|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 3. ATR is required for formation of a subset of UV- and aphidicolin-induced Rad9 foci. (A) HeLa cells were transfected with siRNA oligonucleotides as indicated, exposed to UV and chromatin fractions prepared. Western blot analysis was performed using the indicated antibodies. (B) HeLa cells were transfected with siRNA oligonucleotides as indicated. Cells were treated with UV and harvested for western blotting (upper panel). The percentage of cells exhibiting more than ten Rad9 foci was determined by immunofluorescence (lower panel). *P=0.017 between these two data points indicating statistical significance at the 95% confidence level. (C) Cells were transfected with siRNA and treated as in B. The percentage of cells exhibiting more than ten RPA (p34 subunit) foci was determined by immunofluorescence. (D) HeLa cells were transfected with siRNA oligonucleotides as indicated and 72 hours after transfection were treated with a combination of ATM (10 µM) and DNA-PK (1 µM) inhibitors, or left untreated for 1 hour prior to exposure to aphidicolin. Cells were fractionated to obtain chromatin-enriched proteins and western blotting performed using the indicated antibodies. (E) HeLa cells were transfected with siRNA oligonucleotides as indicated and whole cell extracts prepared for western blotting (upper panel). In parallel, cells were treated with ATM (10 µM) and/or DNA-PK (1 µM) inhibitors, or left untreated prior to exposure to aphidicolin. The percentage of cells exhibiting more than ten Rad9 foci was determined.
|
